A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity - PubMed (original) (raw)
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity
A P Feinberg et al. Anal Biochem. 1983.
Abstract
A technique for conveniently radiolabeling DNA restriction endonuclease fragments to high specific activity is described. DNA fragments are purified from agarose gels directly by ethanol precipitation and are then denatured and labeled with the large fragment of DNA polymerase I, using random oligonucleotides as primers. Over 70% of the precursor triphosphate is routinely incorporated into complementary DNA, and specific activities of over 10(9) dpm/microgram of DNA can be obtained using relatively small amounts of precursor. These "oligolabeled" DNA fragments serve as efficient probes in filter hybridization experiments.
Similar articles
- Direct hybridization of labeled DNA to DNA in agarose gels.
Purrello M, Balazs I. Purrello M, et al. Anal Biochem. 1983 Feb 1;128(2):393-7. doi: 10.1016/0003-2697(83)90391-3. Anal Biochem. 1983. PMID: 6303158 - Discriminatory 32P 3'-end labeling of restriction endonuclease co-digested DNA fragments.
LeBlond GF, Ts'o PO. LeBlond GF, et al. Anal Biochem. 1989 Mar;177(2):413-8. doi: 10.1016/0003-2697(89)90076-6. Anal Biochem. 1989. PMID: 2543232 - Isolation of restriction fragments from large plasmids recovered from bacteria with multiple plasmids.
Via LE, Falkinham JO 3rd. Via LE, et al. Biotechniques. 1991 Oct;11(4):442, 444. Biotechniques. 1991. PMID: 1665338 - Isolation and purification of large DNA restriction fragments from agarose gels.
Moore D, Chory J, Ribaudo RK. Moore D, et al. Curr Protoc Immunol. 2001 May;Chapter 10:Unit 10.5. doi: 10.1002/0471142735.im1005s08. Curr Protoc Immunol. 2001. PMID: 18432696 Review. - Blot hybridisation analysis of genomic DNA.
Vandenplas S, Wiid I, Grobler-Rabie A, Brebner K, Ricketts M, Wållis G, Bester A, Boyd C, Måthew C. Vandenplas S, et al. J Med Genet. 1984 Jun;21(3):164-72. doi: 10.1136/jmg.21.3.164. J Med Genet. 1984. PMID: 6086927 Free PMC article. Review.
Cited by
- Ectopic expression of miRNA172 in tomato (Solanum lycopersicum) reveals novel function in fruit development through regulation of an AP2 transcription factor.
Chung MY, Nath UK, Vrebalov J, Gapper N, Lee JM, Lee DJ, Kim CK, Giovannoni J. Chung MY, et al. BMC Plant Biol. 2020 Jun 19;20(1):283. doi: 10.1186/s12870-020-02489-y. BMC Plant Biol. 2020. PMID: 32560687 Free PMC article. - Isolation and characterization of the complete human beta-myosin heavy chain gene.
Diederich KW, Eisele I, Ried T, Jaenicke T, Lichter P, Vosberg HP. Diederich KW, et al. Hum Genet. 1989 Feb;81(3):214-20. doi: 10.1007/BF00278991. Hum Genet. 1989. PMID: 2522082 - Stable introduction of exogenous DNA into Trypanosoma brucei.
Gibson WC, White TC, Laird PW, Borst P. Gibson WC, et al. EMBO J. 1987 Aug;6(8):2457-61. doi: 10.1002/j.1460-2075.1987.tb02525.x. EMBO J. 1987. PMID: 3665883 Free PMC article. - cDNA cloning and functional expression of the Schistosoma mansoni protective antigen triose-phosphate isomerase.
Shoemaker C, Gross A, Gebremichael A, Harn D. Shoemaker C, et al. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1842-6. doi: 10.1073/pnas.89.5.1842. Proc Natl Acad Sci U S A. 1992. PMID: 1542681 Free PMC article. - Isolation and characterization of expressible cDNA clones encoding the M1 and M2 subunits of mouse ribonucleotide reductase.
Thelander L, Berg P. Thelander L, et al. Mol Cell Biol. 1986 Oct;6(10):3433-42. doi: 10.1128/mcb.6.10.3433-3442.1986. Mol Cell Biol. 1986. PMID: 3025593 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources